Get tips on using MCM4 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - SiHa MCM4
Get tips on using TLR10 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - THP-1 TLR10
Get tips on using TLR10 shRNA (h) Lentiviral Particles to perform shRNA gene silencing Human - THP-1 TLR10
Get tips on using GATA-1 shRNA Plasmids (h) to perform shRNA gene silencing Human - TF‐1 GATA‐1
Get tips on using connexin 43 ShRNA to perform shRNA gene silencing Human - Neuroblastoma cells (SH-SY5Y) Connexin 43 lentiviral particles
Get tips on using pLX-sgRNA to perform CRISPR Human - Deletion CIITA
Get tips on using Cas9 sgRNA vector to perform CRISPR Human - Activation ESR1
Is a knockdown using shRNA permanent and if not is there a known duration?
Get tips on using peSpCas9(1.1)-2×sgRNA (empty, donor) to perform CRISPR Human - Repression SLC7A11
The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.
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